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Article: Bacterial and Viral Pathogens with One Health Relevance in Invasive Raccoons (

Reinhardt, Nico P / Köster, Judith / Thomas, Astrid / Arnold, Janosch / Fux, Robert / Straubinger, Reinhard K

2023 Volume 12, Issue 3

Abstract: In Europe, raccoons are invasive neozoons with their largest population in Germany. Globally, this mesocarnivore acts as a wildlife reservoir for many (non-)zoonotic (re-)emerging pathogens, but very little epidemiological data is available for southwest ...

Abstract	In Europe, raccoons are invasive neozoons with their largest population in Germany. Globally, this mesocarnivore acts as a wildlife reservoir for many (non-)zoonotic (re-)emerging pathogens, but very little epidemiological data is available for southwest Germany. This exploratory study aimed to screen free-ranging raccoons in Baden-Wuerttemberg (BW, Germany) for the occurrence of selected pathogens with One Health relevance. Organ tissue and blood samples collected from 102 animals, obtained by hunters in 2019 and 2020, were subsequently analysed for two bacterial and four viral pathogens using a qPCR approach. Single samples were positive for the carnivore protoparvovirus-1 (7.8%, n = 8), canine distemper virus (6.9%, n = 7), pathogenic
Language	English
Publishing date	2023-03-01
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2695572-6
ISSN	2076-0817
ISSN	2076-0817
DOI	10.3390/pathogens12030389
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Article ; Online: Prevalence of Taylorella equigenitalis in an Icelandic horse population

Solbach, Veronika / Grabatin, Markus / Fux, Robert / Zablotski, Yury / Zerbe, Holm / Goehring, Lutz S / Witte, Tanja S

Journal of Equine Veterinary Science. 2023 June, v. 125 p.104749-

2023

Abstract: Contagious Equine Metritis (CEM) is an infectious disease caused by Taylorella equigenitalis (TEq). In mares, clinical as well as subclinical infections have the potential to cause short-term infertility. Stallions are generally believed to be ... ...

Abstract	Contagious Equine Metritis (CEM) is an infectious disease caused by Taylorella equigenitalis (TEq). In mares, clinical as well as subclinical infections have the potential to cause short-term infertility. Stallions are generally believed to be asymptomatic carrier of TEq. Transmission commonly occurs venereally and therefore poses a risk for horses in less controlled natural mating programs (Timoney, Comp. Immun. Microbiol. Infect. Dis. 1996, 3: 199-204). In the Icelandic breed, pasture breeding is routinely performed. Therefore, the aim of this study was to investigate the prevalence of TEq in intact stallions, mares and geldings of the Icelandic breed. We hypothesized that there is a high prevalence of TEq in Icelandic horses independent of their breeding purpose. In total, 437 Icelandic horses from 16 randomly selected farms in southern Germany and Austria were examined. In a first trial, 76 intact Icelandic males from 10 farms were compared to a reference group (n=86) consisting of 35 Haflinger and 51 Draft intact male horses from 31 farms. Breeds of the reference group are also known to be used in natural mating programs. In a second trial, 105 geldings and 256 mares from 11 farms were tested for presence of TEq. In both trials, swabs from standardized localizations of the male (penis) or female (clitoris) genital tract were collected (WOAH for TEq). The presence of TEq DNA was determined using quantitativePCR. There was a significant higher prevalence of TEq-positive samples in Icelandic intact males (n=23) compared to the reference group (n=4; p=0.0001). Both groups were further differentiated regarding their actual use for breeding. Pairwise comparison of both groups resulted in a significantly lower number of TEq-positive intact males used for breeding compared to the horses without breeding (reference group: p=0.019; Icelandic group: p=0.006). In the second trial, a higher prevalence of TEq-positive geldings compared to mares (36.2% and 5.5%, respectively) was detected. Mares were further differentiated in mares with breeding history (n=134) and mares without (n=122). A higher prevalence in mares without breeding history could be found (2.2% and 9.0%, respectively). The hypothesis could be confirmed that in the Icelandic breed a higher prevalence of TEq-positive stallions compared to stallions also used in natural mating programs exists. Furthermore, TEq has a higher prevalence in geldings compared to mares of Icelandic horses. Therefore, geldings might represent a reservoir of TEq in this breed. The spread of CEM is of economic importance and should be considered even in horses never used for breeding. Further studies investigating the genotype of the CEM strains in the Icelandic breed might help to understand further ways of transmission.
Keywords	DNA ; Haflinger ; Icelandic (horse breed) ; Taylorella equigenitalis ; carrier state ; contagious equine metritis ; females ; genotype ; horses ; males ; pastures ; penis ; risk ; veterinary medicine ; Austria ; Germany
Language	English
Dates of publication	2023-06
Publishing place	Elsevier Inc.
Document type	Article ; Online
ZDB-ID	2102631-2
ISSN	1542-7412 ; 0737-0806
ISSN (online)	1542-7412
ISSN	0737-0806
DOI	10.1016/j.jevs.2023.104749
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Article: Detection of porcine circovirus type 3 DNA in serum and semen samples of boars from a German boar stud

Eddicks, Matthias / Müller, Michael / Fux, Robert / Ritzmann, Mathias / Stadler, Julia

veterinary journal. 2022 Jan., v. 279

2022

Abstract: Porcine circovirus type 3 (PCV3) is regularly reported in association with various clinical presentations, including porcine dermatitis and nephropathy syndrome (PDNS)-like lesions, respiratory signs, congenital tremor, and reproductive disorders. To ... ...

Abstract	Porcine circovirus type 3 (PCV3) is regularly reported in association with various clinical presentations, including porcine dermatitis and nephropathy syndrome (PDNS)-like lesions, respiratory signs, congenital tremor, and reproductive disorders. To investigate the epidemiology of PCV3 in a boar stud, we analysed fresh boar semen and matching sera from 181 boars from a German stud supplying semen for artificial insemination (AI) to approximately 740 breeder farms for PCV3 DNA. PCV3 DNA was detected in 1.7% semen samples and 24.3% sera. Spearman rho correlation demonstrated a significant positive correlation between boar age and quantitative DNA (by PCR quantification cycles [Cq] values) in serum samples (r = 0.636; P < 0.001). Sera from boars up to 12 months of age had higher viral loads (P < 0.001) and were PCV3-positive significantly more often (P < 0.01) than older boars. Detection of PCV3 DNA was not associated with breed (P> 0.05). PCV3 DNA was detected sporadically in fresh boar semen. Based on the assumption that processing fresh semen reduces viral load in semen used for AI, it is likely that the risk of sexual transmission of PCV3 during AI in is low. However, young boars may contribute to the maintenance of PCV3 infection in boar studs.
Keywords	DNA ; Porcine circovirus-3 ; artificial insemination ; blood serum ; boars ; dermatitis ; kidney diseases ; risk ; semen ; viral load
Language	English
Dates of publication	2022-01
Publishing place	Elsevier Ltd
Document type	Article
ZDB-ID	428614-5
ISSN	1532-2971 ; 0372-5545 ; 1090-0233
ISSN (online)	1532-2971
ISSN	0372-5545 ; 1090-0233
DOI	10.1016/j.tvjl.2021.105784
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Article ; Online: Detection of porcine circovirus type 3 DNA in serum and semen samples of boars from a German boar stud.

Eddicks, Matthias / Müller, Michael / Fux, Robert / Ritzmann, Mathias / Stadler, Julia

Veterinary journal (London, England : 1997)

2021 Volume 279, Page(s) 105784

Abstract	Porcine circovirus type 3 (PCV3) is regularly reported in association with various clinical presentations, including porcine dermatitis and nephropathy syndrome (PDNS)-like lesions, respiratory signs, congenital tremor, and reproductive disorders. To investigate the epidemiology of PCV3 in a boar stud, we analysed fresh boar semen and matching sera from 181 boars from a German stud supplying semen for artificial insemination (AI) to approximately 740 breeder farms for PCV3 DNA. PCV3 DNA was detected in 1.7% semen samples and 24.3% sera. Spearman rho correlation demonstrated a significant positive correlation between boar age and quantitative DNA (by PCR quantification cycles [Cq] values) in serum samples (r = 0.636; P < 0.001). Sera from boars up to 12 months of age had higher viral loads (P < 0.001) and were PCV3-positive significantly more often (P < 0.01) than older boars. Detection of PCV3 DNA was not associated with breed (P> 0.05). PCV3 DNA was detected sporadically in fresh boar semen. Based on the assumption that processing fresh semen reduces viral load in semen used for AI, it is likely that the risk of sexual transmission of PCV3 during AI in is low. However, young boars may contribute to the maintenance of PCV3 infection in boar studs.
MeSH term(s)	Animals ; Circoviridae Infections/epidemiology ; Circoviridae Infections/veterinary ; Circovirus/genetics ; Insemination, Artificial/veterinary ; Male ; Semen ; Swine ; Swine Diseases/epidemiology
Language	English
Publishing date	2021-12-10
Publishing country	England
Document type	Journal Article
ZDB-ID	428614-5
ISSN	1532-2971 ; 0372-5545 ; 1090-0233
ISSN (online)	1532-2971
ISSN	0372-5545 ; 1090-0233
DOI	10.1016/j.tvjl.2021.105784
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Article: Identification and Characterization of Circular Single-Stranded DNA Genomes in Sheep and Goat Milk

König, Marie-Thérèse / Fux, Robert / Link, Ellen / Sutter, Gerd / Märtlbauer, Erwin / Didier, Andrea

Viruses. 2021 Oct. 28, v. 13, no. 11

2021

Abstract: In recent years, a variety of circular replicase-encoding single-stranded (CRESS) DNA viruses and unclassified virus-like DNA elements have been discovered in a broad range of animal species and environmental samples. Key questions to be answered concern ...

Abstract	In recent years, a variety of circular replicase-encoding single-stranded (CRESS) DNA viruses and unclassified virus-like DNA elements have been discovered in a broad range of animal species and environmental samples. Key questions to be answered concern their presence in the human diet and their potential impact on disease emergence. Especially DNA elements termed bovine meat and milk factors (BMMF) are suspected to act as co-factors in the development of colon and breast cancer. To expand our knowledge on the occurrence of these potential pathogens in human nutrition, a total of 73 sheep and 40 goat milk samples were assayed by combining rolling circle amplification (RCA), PCR and Sanger sequencing. The present study further includes retail milk from the aforementioned species. We recovered 15 single stranded (ss) circular genomes. Of those, nine belong to the family Genomoviridae and six are members of the unclassified group of BMMF. Thus, dairy sheep and goats add to dispersal of CRESS viruses and circular ssDNA elements, which enter the food chain via milk. The presence of these entities is therefore more widespread in Bovidae than initially assumed and seems to be part of the common human nutrition.
Keywords	beef ; breast neoplasms ; circular DNA ; colon ; dairy sheep ; food chain ; genome ; goat milk ; human nutrition ; milk ; sheep
Language	English
Dates of publication	2021-1028
Publishing place	Multidisciplinary Digital Publishing Institute
Document type	Article
ZDB-ID	2516098-9
ISSN	1999-4915
ISSN	1999-4915
DOI	10.3390/v13112176
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Article: Circular Rep-Encoding Single-Stranded DNA Sequences in Milk from Water Buffaloes (Bubalus arnee f. bubalis)

König, Marie-T. / Fux, Robert / Link, Ellen / Sutter, Gerd / Märtlbauer, Erwin / Didier, Andrea

Viruses. 2021 June 07, v. 13, no. 6

2021

Abstract: Isolation and characterization of circular replicase-encoding single-stranded (ss) DNA from animal, plant and environmental samples are rapidly evolving in virology. We detected 21 circular DNA elements, including one genomoviral sequence, in individual ... ...

Abstract	Isolation and characterization of circular replicase-encoding single-stranded (ss) DNA from animal, plant and environmental samples are rapidly evolving in virology. We detected 21 circular DNA elements, including one genomoviral sequence, in individual milk samples from domesticated Asian water buffaloes (Bubalus arnee f. bubalis). Most of the obtained genomes are related to Sphinx 1.76 and Sphinx 2.36 sequences and share a high degree of similarity to recently published circular DNAs—named BMMF (bovine meat and milk factors)—that have been isolated from commercial milk, as well as from bovine serum. Characteristic features such as rep genes, tandem repeats and inverted repeats were detected. These BMMF have recently been found to be present in taurine-type dairy cattle breeds descending from the aurochs (Bos primigenius). Importantly, the occurrence of BMMF has been linked to the higher incidence of colorectal and breast cancer in North America and Western Europe compared with Asia. This is the first report of circular ssDNA detected in milk from the domesticated form of the wild Asian water buffalo (B. arnee) belonging to the subfamily Bovinae. This novelty should be taken into account in view of the above-mentioned cancer hypothesis.
Keywords	Bubalus bubalis arnee ; Sphinx ; beef ; blood serum ; breast neoplasms ; buffaloes ; circular DNA ; dairy cattle ; milk ; single-stranded DNA ; virology ; Asia ; North America ; Western European region
Language	English
Dates of publication	2021-0607
Publishing place	Multidisciplinary Digital Publishing Institute
Document type	Article
ZDB-ID	2516098-9
ISSN	1999-4915
ISSN	1999-4915
DOI	10.3390/v13061088
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Article ; Online: Discriminating the eight genotypes of the porcine circovirus type 2 with TaqMan-based real-time PCR.

Link, Ellen Kathrin / Eddicks, Matthias / Nan, Liangliang / Ritzmann, Mathias / Sutter, Gerd / Fux, Robert

Virology journal

2021 Volume 18, Issue 1, Page(s) 70

Abstract: Background: The porcine circovirus type 2 (PCV2) is divided into eight genotypes including the previously described genotypes PCV2a to PCV2f and the two new genotypes PCV2g and PCV2h. PCV2 genotyping has become an important task in molecular ... ...

Abstract	Background: The porcine circovirus type 2 (PCV2) is divided into eight genotypes including the previously described genotypes PCV2a to PCV2f and the two new genotypes PCV2g and PCV2h. PCV2 genotyping has become an important task in molecular epidemiology and to advance research on the prophylaxis and pathogenesis of PCV2 associated diseases. Standard genotyping of PCV2 is based on the sequencing of the viral genome or at least of the open reading frame 2. Although, the circovirus genome is small, classical sequencing is time consuming, expensive, less sensitive and less compatible with mass testing compared with modern real-time PCR assays. Here we report about a new PCV2 genotyping method using qPCR. Methods: Based on the analysis of several hundred PCV2 full genome sequences, we identified PCV2 genotype specific sequences or single-nucleotide polymorphisms. We designed six TaqMan PCR assays that are specific for single genotypes PCV2a to PCV2f and two qPCRs targeting two genotypes simultaneously (PCV2g/PCV2d and PCV2h/PCV2c). To improve specific binding of oligonucleotide primers and TaqMan probes, we used locked nucleic acid technology. We evaluated amplification efficiency, diagnostic sensitivity and tested assay specificity for the respective genotypes. Results: All eight PCV2 genotype specific qPCRs demonstrated appropriate amplification efficiencies between 91 and 97%. Testing samples from an epidemiological field study demonstrated a diagnostic sensitivity of the respective genotype specific qPCR that was comparable to a highly sensitive pan-PCV2 qPCR system. Genotype specificity of most qPCRs was excellent. Limited unspecific signals were obtained when a high viral load of PCV2b was tested with qPCRs targeting PCV2d or PCV2g. The same was true for the PCV2a specific qPCR when high copy numbers of PCV2d were tested. The qPCR targeting PCV2h/PCV2c showed some minor cross-reaction with PCV2d, PCV2f and PCV2g. Conclusion: Genotyping of PCV2 is important for routine diagnosis as well as for epidemiological studies. The introduced genotyping qPCR system is ideal for mass testing and should be a valuable complement to PCV2 sequencing, especially in the case of simultaneous infections with multiple PCV2 genotypes, subclinically infected animals or research studies that require large sample numbers.
MeSH term(s)	Animals ; Circoviridae Infections/diagnosis ; Circoviridae Infections/veterinary ; Circovirus/genetics ; Genotype ; Phylogeny ; Real-Time Polymerase Chain Reaction ; Swine ; Swine Diseases/diagnosis ; Swine Diseases/virology
Language	English
Publishing date	2021-04-07
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2160640-7
ISSN	1743-422X ; 1743-422X
ISSN (online)	1743-422X
ISSN	1743-422X
DOI	10.1186/s12985-021-01541-z
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Article ; Online: Identification and Characterization of Circular Single-Stranded DNA Genomes in Sheep and Goat Milk.

König, Marie-Thérèse / Fux, Robert / Link, Ellen / Sutter, Gerd / Märtlbauer, Erwin / Didier, Andrea

Viruses

2021 Volume 13, Issue 11

Abstract	In recent years, a variety of circular replicase-encoding single-stranded (CRESS) DNA viruses and unclassified virus-like DNA elements have been discovered in a broad range of animal species and environmental samples. Key questions to be answered concern their presence in the human diet and their potential impact on disease emergence. Especially DNA elements termed bovine meat and milk factors (BMMF) are suspected to act as co-factors in the development of colon and breast cancer. To expand our knowledge on the occurrence of these potential pathogens in human nutrition, a total of 73 sheep and 40 goat milk samples were assayed by combining rolling circle amplification (RCA), PCR and Sanger sequencing. The present study further includes retail milk from the aforementioned species. We recovered 15 single stranded (ss) circular genomes. Of those, nine belong to the family
MeSH term(s)	Animals ; Cattle ; DNA Viruses/classification ; DNA Viruses/genetics ; DNA, Circular/isolation & purification ; DNA, Single-Stranded/isolation & purification ; DNA, Viral/isolation & purification ; Genome, Viral ; Germany ; Goats ; Milk/virology ; Phylogeny ; Polymerase Chain Reaction ; Sheep
Chemical Substances	DNA, Circular ; DNA, Single-Stranded ; DNA, Viral
Language	English
Publishing date	2021-10-28
Publishing country	Switzerland
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2516098-9
ISSN	1999-4915 ; 1999-4915
ISSN (online)	1999-4915
ISSN	1999-4915
DOI	10.3390/v13112176
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Article: Activation of interferon regulatory factor 3 by replication-competent vaccinia viruses improves antitumor efficacy mediated by T cell responses.

Riederer, Stephanie / Fux, Robert / Lehmann, Michael H / Volz, Asisa / Sutter, Gerd / Rojas, Juan J

Molecular therapy oncolytics

2021 Volume 22, Page(s) 399–409

Abstract: Recently, oncolytic vaccinia viruses (VACVs) have shown their potential to provide for clinically effective cancer treatments. The reason for this clinical usefulness is not only the direct destruction of infected cancer cells but also activation of ... ...

Abstract	Recently, oncolytic vaccinia viruses (VACVs) have shown their potential to provide for clinically effective cancer treatments. The reason for this clinical usefulness is not only the direct destruction of infected cancer cells but also activation of immune responses directed against tumor antigens. For eliciting a robust antitumor immunity, a dominant T helper 1 (Th1) cell differentiation of the response is preferred, and such polarization can be achieved by activating the Toll-like receptor 3 (TLR3)-interferon regulatory factor 3 (IRF3) signaling pathway. However, current VACVs used as oncolytic viruses to date still encode several immune evasion proteins involved in the inhibition of this signaling pathway. By inactivating genes of selected regulatory virus proteins, we aimed for a candidate virus with increased potency to activate cellular antitumor immunity but at the same time with a fully maintained replicative capacity in cancer cells. The removal of up to three key genes (
Language	English
Publishing date	2021-06-04
Publishing country	United States
Document type	Journal Article
ISSN	2372-7705
ISSN	2372-7705
DOI	10.1016/j.omto.2021.06.001
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Article ; Online: Circular Rep-Encoding Single-Stranded DNA Sequences in Milk from Water Buffaloes (

König, Marie-T / Fux, Robert / Link, Ellen / Sutter, Gerd / Märtlbauer, Erwin / Didier, Andrea

Viruses

2021 Volume 13, Issue 6

Abstract	Isolation and characterization of circular replicase-encoding single-stranded (ss) DNA from animal, plant and environmental samples are rapidly evolving in virology. We detected 21 circular DNA elements, including one genomoviral sequence, in individual milk samples from domesticated Asian water buffaloes (
MeSH term(s)	Animals ; Base Sequence ; Biomarkers ; Buffaloes/genetics ; Cattle ; Cloning, Molecular ; DNA Viruses/genetics ; DNA, Circular ; DNA, Single-Stranded ; Milk/chemistry ; Phylogeny ; Polymerase Chain Reaction ; Sequence Analysis, DNA
Chemical Substances	Biomarkers ; DNA, Circular ; DNA, Single-Stranded
Language	English
Publishing date	2021-06-07
Publishing country	Switzerland
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2516098-9
ISSN	1999-4915 ; 1999-4915
ISSN (online)	1999-4915
ISSN	1999-4915
DOI	10.3390/v13061088
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